PurposeCas9 expression controlled by the nmt41 promoter
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||121145||Standard format: Plasmid sent in bacteria as agar stab||1||$75|
This material is available to academics and nonprofits only.
- Backbone size w/o insert (bp) 8460
- Total vector size (bp) 12736
Vector typeYeast Expression, CRISPR
Selectable markersS.pombe ura4
Growth in Bacteria
Copy numberHigh Copy
Gene/Insert namehumanized Streptococcus pyogenes Cas9
Insert Size (bp)4276
- Promoter nmt41
/ Fusion Protein
- Flag (N terminal on insert)
- Cloning method Restriction Enzyme
- 5′ cloning site Xho I (destroyed during cloning)
- 3′ cloning site BglII (not destroyed)
- 5′ sequencing primer TCTCACTTTCTGACTTATAGTCGCT
- 3′ sequencing primer AGCAGTACTGGCAAGGGAGAC (Common Sequencing Primers)
Ran, F.A., Hsu, P.D.P., Wright, J., Agarwala, V., Scott, D. a and Zhang, F. (2013) Genome engineering using the CRISPR-Cas9 system. Nat. Protoc., 8, 2281–2308. After prepping the DNA, it is recommended to heat the DNA to 65C before digesting with Bbs I. Supplementary data is available at Figshare: https://doi.org/10.25387/g3.7685642.
These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.
For your Materials & Methods section:pAH235 was a gift from Katsunori Tanaka (Addgene plasmid # 121145 ; http://n2t.net/addgene:121145 ; RRID:Addgene_121145)
For your References section:Short-Homology-Mediated CRISPR/Cas9-Based Method for Genome Editing in Fission Yeast. Hayashi A, Tanaka K. G3 (Bethesda). 2019 Feb 12. pii: g3.118.200976. doi: 10.1534/g3.118.200976. 10.1534/g3.118.200976 PubMed 30755408